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- Title
Assessment of small ligand-protein interactions by electrophoretic mobility shift assay using DNA-modified ligand as a sensing probe.
- Authors
Funabashi, Hisakage; Ubukata, Michito; Ebihara, Takashi; Aizawa, Masuo; Mie, Masayasu; Kobatake, Eiry
- Abstract
The interaction between a small ligand and a protein were assessed by electrophoretic mobility shift assay. A sensing probe was created by modifying the model ligand, biotin, with DNA. The complex of DNA-modified ligand and anti-biotin antibody or streptavidin as a target protein was analyzed by agarose gel electrophoresis. The band corresponding to the DNA-modified ligand was shifted in the presence of the target protein, and the intensities of the shifted bands were decreased by adding increasing concentrations of free ligand ranging from 0.1 μM to 100 μM. From this calibration the concentration of ligand in the samples could be determined, allowing for evaluation of the interaction between a small ligand and its target.
- Subjects
LIGANDS (Biochemistry); PROTEINS; GEL electrophoresis; BACTERIAL proteins; BIOTIN
- Publication
Biotechnology Letters, 2007, Vol 29, Issue 5, p785
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1007/s10529-006-9301-6